The Cell Counting Kit-8 (CCK8, Meilunbio, Dalian, China, MA0218) and 5-ethynyl-2′-deoxyuridine (EdU, Meilunbio, MA0425) labeling assay were used to measure cell proliferation based on the manufacturer’s protocols. In brief, for the CCK8 assay, HA and A172 cells were seeded inside the 96-well microplates and cultured overnight. After incubation with MitoQ for 2 h, the cells were radiated using X-rays at doses of 4 Gy or carbon ion at 2 Gy. Then, 10 μL of CCK-8 reagent was added to each well after 24 h and incubated for 2 h. The optical density (OD) was determined at 450 nm through a multi-plate reader (Tecan Infinite M200, Männedorf, Switzerland).
For the EdU labeling assay, HA and A172 cells were seeded into a 35 mm confocal dish and cultured overnight. After treatment using MitoQ for 2 h, the cells were radiated with 4 Gy X-rays or 2 Gy carbon ion. Based on the manufacturer’s instructions, the EdU kit was utilized for assessing cell proliferative ability after irradiating for 24 h. Hoechst 33,342 (nuclear staining) was utilized to counterstain cells. The images were captured with Laser confocal microscopy.
For the EdU labeling assay, HA and A172 cells were seeded into a 35 mm confocal dish and cultured overnight. After treatment using MitoQ for 2 h, the cells were radiated with 4 Gy X-rays or 2 Gy carbon ion. Based on the manufacturer’s instructions, the EdU kit was utilized for assessing cell proliferative ability after irradiating for 24 h. Hoechst 33,342 (nuclear staining) was utilized to counterstain cells. The images were captured with Laser confocal microscopy.
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